Cytotoxic and Antitumor Activity of Lactaptin in Combination with Autophagy Inducers and Inhibitors
Cytotoxic and Antitumor Activity of Lactaptin in Combination with Autophagy Inducers and Inhibitors
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DOI：
10.1155/2019/4087160
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发表时间：
2019-06-17
影响因子：
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通讯作者：
Koval, Olga A.
中科院分区：
文献类型：
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作者：
Bagamanshina, Anastasia V.;Troitskaya, Olga S.;Koval, Olga A.
自噬是一种降解过程，其中细胞器和蛋白质被回收以恢复体内平衡和细胞代谢。自噬可以是促生存或促死亡过程，并且仍然是细胞活力最基本的过程之一。因此，自噬调节是强化抗癌治疗的重要方法。早些时候，我们已经证明，人乳蛋白乳肽 (RL2) 的重组类似物可诱导各种培养的癌细胞凋亡，并激活微管相关蛋白 1 轻链 3 (LC3) 的脂化。在本研究中，我们研究了自噬抑制剂 - 氯喹 (CQ)、Ku55933 (Ku) 和 3-甲基腺嘌呤 (3MA) - 或诱导剂 - 雷帕霉素 (Rap) - 是否可以增强乳酶类似物在癌细胞中的细胞毒活性及其在小鼠模型中的抗癌活性。 Western Blot 分析显示，RL2 在培养早期阶段诱导 MDA-MB-231 和 MCF-7 细胞中的短期自噬，并且这些数据通过自噬体/自噬溶酶体形成的透射电子显微镜得到证实。 RL2 刺激活性氧 (ROS) 产生、自噬体积累、通过 LC3I 加工成 LC3II 上调 ATG5，以及下调 p62/sequestosome 1 (p62)。我们已经证明自噬调节剂 CQ、Ku 和 Rap 可以协同增加 RL2 的细胞毒性，并且 RL2 与 CQ 一起诱导自噬细胞死亡。此外，CQ、Ku 和 Rap 与 RL2 组合可降低溶酶体蛋白酶组织蛋白酶 D 的活性。更重要的是，将 RL2 与 CQ 组合，我们提高了小鼠的抗肿瘤效果。检测到两种类型的自噬调节剂、抑制剂和诱导剂与 RL2 对癌细胞的协同细胞毒性作用使我们相信这些组合可以成为新抗癌方法的基础。最后，我们假设 CQ 和 Rap 促进短期 RL2 诱导的自噬与最终的自噬细胞死亡相互关联。
Autophagy is a degradative process in which cellular organelles and proteins are recycled to restore homeostasis and cellular metabolism. Autophagy can be either a prosurvival or a prodeath process and remains one of the most fundamental processes for cell vitality. Thus autophagy modulation is an important approach for reinforcement anticancer therapeutics. Earlier we have demonstrated that recombinant analog of human milk protein lactaptin (RL2) induced apoptosis of various cultured cancer cells and activated lipidation of microtubule-associated protein 1 light chain 3 (LC3). In this study we investigated whether autophagy inhibitors-chloroquine (CQ), Ku55933 (Ku), and 3-methyladenine (3MA)-or inducer-rapamycin (Rap)-can enhance cytotoxic activity of lactaptin analog in cancer cells and its anticancer activity in the mice model. Western Blot analysis revealed that RL2 induced short-term autophagy in MDA-MB-231 and MCF-7 cells at early stages of incubation and that these data were confirmed by the transmission electron microscopy of autophagosome/autophagolysosome formation. RL2 stimulates reactive oxygen species (ROS) production, autophagosomes accumulation, upregulation of ATG5 with processing of LC3I to LC3II, and downregulation of p62/sequestosome 1 (p62). We have shown that autophagy modulators, CQ, Ku, and Rap, synergistically increased cytotoxicity of RL2, and RL2 with CQ induced autophagic cell death. In addition, CQ, Ku, and Rap in combination with RL2 decreased activity of lysosomal protease Cathepsin D. More importantly, combining RL2 with CQ, we improved antitumor effect in mice. Detected synergistic cytotoxic effects of both types of autophagy regulators, inhibitors, and inducers with RL2 against cancer cells allow us to believe that these combinations can be a basis for the new anticancer approach. Finally, we suppose that CQ and Rap promoting of short-term RL2-induced autophagy interlinks with final autophagic cell death.